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Biochemical and genetic analyses of the U5, U6, and U4/U6 x U5 small nuclear ribonucleoproteins from Saccharomyces cerevisiae

Stevens, Scott W. and Barta, Imre and Ge, Helen Y. and Moore, Roger E. and Young, Mary K. and Lee, Terry D. and Abelson, John (2001) Biochemical and genetic analyses of the U5, U6, and U4/U6 x U5 small nuclear ribonucleoproteins from Saccharomyces cerevisiae. RNA, 7 (11). pp. 1543-1553. ISSN 1355-8382.

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We have purified the yeast U5 and U6 pre-mRNA splicing small nuclear ribonucleoproteins (snRNPs) by affinity chromatography and analyzed the associated polypeptides by mass spectrometry. The yeast U5 snRNP is composed of the two variants of U5 snRNA, six U5-specific proteins and the 7 proteins of the canonical Sm core. The U6 snRNP is composed of the U6 snRNA, Prp24, and the 7 Sm-Like (LSM) proteins. Surprisingly, the yeast DEAD-box helicase-like protein Prp28 is stably associated with the U5 snRNP, yet is absent from the purified U4/U6.U5 snRNR A novel yeast U5 and four novel yeast U4/U6.U5 snRNP polypeptides were characterized by genetic and biochemical means to demonstrate their involvement in the pre-mRNA splicing reaction. We also show that, unlike the human tri-snRNP, the yeast tri-snRNP dissociated upon addition of ATP or dATP.

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Additional Information:Copyright © 2001 by RNA Society. Received July 3, 2001; returned for revision August 14, 2001; revised manuscript received August 27, 2001. We gratefully acknowledge the generous gift of strains and hybridoma cells from Christine Guthrie. We are also grateful for helpful conversations with the members of the Abelson, Guthrie, and Lührmann laboratories and to Alexander Gottschalk and Patrizia Fabrizio for communicating results prior to publication. This work was supported by National Institutes of Health Grants GM32627 to J.A., and CA33572 to T.D.L. and by an American Cancer Society postdoctoral fellowship PF4447 to S.W.S.
Subject Keywords:affinity chromatography; mass spectrometry; spliceosome assembly
Issue or Number:11
Record Number:CaltechAUTHORS:STErna01
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:2986
Deposited By: Tony Diaz
Deposited On:10 May 2006
Last Modified:02 Oct 2019 22:58

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